PMID:21917918

Citation	Agarwal, S, Agarwal, S, Pancholi, P and Pancholi, V (2011) Role of serine/threonine phosphatase (SP-STP) in Streptococcus pyogenes physiology and virulence. J. Biol. Chem. 286:41368-80
Abstract	Reversible phosphorylation is the key mechanism regulating several cellular events in prokaryotes and eukaryotes. In prokaryotes, signal transduction is perceived to occur primarily via the two-component signaling system involving histidine kinases and cognate response regulators. Although an alternative regulatory pathway controlled by the eukaryote-type serine/threonine kinase (Streptococcus pyogenes serine/threonine kinase; SP-STK) has been shown to modulate bacterial growth, division, adherence, invasion, and virulence in group A Streptococcus (GAS; S. pyogenes), the precise role of the co-transcribing serine/threonine phosphatase (SP-STP) has remained enigmatic. In this context, this is the first report describing the construction and characterization of non-polar SP-STP mutants in two different strains of Type M1 GAS. The STP knock-out mutants displayed increased bacterial chain lengths in conjunction with thickened cell walls, significantly reduced capsule and hemolysin production, and restoration of the phenotypes postcomplementation. The present study also reveals important contribution of cognately regulated-reversible phosphorylation by SP-STK/SP-STP on two major response regulators of two-component systems, WalRK and CovRS. We also demonstrate a distinct role of SP-STP in terms of expression of surface proteins and SpeB in a strain-specific manner. Further, the attenuation of virulence in the absence of STP and its restoration only in the complemented strains that were generated by the use of a low copy plasmid and not by a high copy one emphasize not only the essential role of STP in virulence but also highlight the tightly regulated SP-STP/SP-STK-mediated cognate functions. SP-STP thus is an important regulator of GAS virulence and plays a critical role in GAS pathogenesis.
Links	PubMed PMC3308849 Online version:10.1074/jbc.M111.286690
Keywords	Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Cell Line; Gene Expression Regulation, Bacterial/physiology; Humans; Mutation; Phosphoprotein Phosphatases/genetics; Phosphoprotein Phosphatases/metabolism; Phosphorylation/physiology; Streptococcus pyogenes/enzymology; Streptococcus pyogenes/pathogenicity; Virulence Factors/genetics; Virulence Factors/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
STRPY:A0A2W5AMW9		GO:0018107: peptidyl-threonine phosphorylation	ECO:0000314:		P		Figure 3C and Figure 3D STK (Serine/threonine-protein kinase PrkC) STK (Stk1 family PASTA domain-containing Ser/Thr kinase) (change to this in notes) Streptococcus pyogenes “The in vitro phosphorylation assay revealed that SP-STK was able to phosphorylate SP-WalR (Fig. 3C) specifically at threonine residues, as is evident by TLC (Fig. 3D).”	complete CACAO 13378
STRP1:Q99YM9		GO:0035970: peptidyl-threonine dephosphorylation	ECO:0000314:		P		Figure 3C STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “The SP-STP was able to dephosphorylate the SP-STK-phosphorylated SP-WalR (Fig. 3C), demonstrating that this couple functions as a cognate kinase-phosphatase pair.”	complete CACAO 13336
STRP1:Q99YM9		GO:0045893: positive regulation of transcription, DNA-templated	ECO:0000315:		P		Figure 3A and Figure 3B STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “In support of this, we also observed a simultaneous up-regulation of walR (2.7–3.2-fold) transcript at an early time point in both of the strains (Fig. 3, table below A and B).”	complete CACAO 13337
STRP1:Q99YM9		GO:0019836: hemolysis by symbiont of host erythrocytes	ECO:0000315:		P		Figure 4B STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “hemolysin production in the wild-type M1SF370 and M1T1 and their corresponding mutant and complemented strains. Complete lysis of shRBCs achieved upon the addition of 1% Triton X-100 was taken as control (100%).”	complete CACAO 13351
STRP1:Q99YM9		GO:1905228: positive regulation of adhesion of symbiont to host epithelial cell	ECO:0000315:		P		Figure 5A STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “The ability of the mutants and stp-complemented strains to adhere to...Detroit 562 human pharyngeal cells in comparison with that of the wild-type strains was determined by calculating adherence (percentage of initial inoculum)”	complete CACAO 13353
STRP1:Q99YM9		GO:0000918: division septum site selection	ECO:0000315:		P		Figure 2C and Figure 2D STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “Similar defects observed by transmission electron microscopy (Fig. 2, C and D) in conjunction with relatively thicker cell walls indicated that SP-STP plays a role in the regulation of septa formation and in turn cell division in GAS."	complete CACAO 13357
STRP1:Q99YM9		GO:0045717: negative regulation of fatty acid biosynthetic process	ECO:0000315:		P		Table 1 STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “The total lipid content analysis (fatty acid methyl ester) of the wild type and its corresponding STP mutant revealed that both saturated fatty acid (SFA) and unsaturated fatty acid (UFA) content in the mutant were 2–3-fold up-regulated”	complete CACAO 13379
STRP1:Q99YM9		GO:0075294: positive regulation by symbiont of entry into host	ECO:0000315:		P		Figure 5B STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “The ability of the mutants and stp-complemented strains to adhere to...Detroit 562 human pharyngeal cells in comparison with that of the wild-type strains was determined by calculating...invasion (percentage of adherence) (B)”	complete CACAO 13380
STRP1:Q99YM9		GO:0062085: positive regulation of capsule polysaccharide biosynthetic process	ECO:0000315:		P		Figure 4a STP (Putative phosphoprotein phosphatase) Streptococcus pyogenes “the comparison of capsule content based on the biochemical estimation of the hyaluronic acid for the STP mutants and complemented GAS strains with respect to their corresponding wild-type strains.”	complete CACAO 13384
STRPY:A0A2X4C3R8		GO:0018107: peptidyl-threonine phosphorylation	IDA: Inferred from Direct Assay:	P	Figure 3C and Figure 3D STK (Serine/threonine-protein kinase PrkC) Streptococcus pyogenes “The in vitro phosphorylation assay revealed that SP-STK was able to phosphorylate SP-WalR (Fig. 3C) specifically at threonine residues, as is evident by TLC (Fig. 3D).”	complete CACAO 13338

References

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